In search for globally disordered apo-parvalbumins: Case of parvalbumin β-1 from coho salmon

Parvalbumin (PA) is a classical EF-hand calcium-binding protein of muscle, neuronal, and other tissues, and a major fish allergen. Although certain apo-PAs lack tertiary structure, functional implications of that feature and its structural prerequisites remain unclear. In a search for unstable PM, we probed conformational stability of parvalbumin beta-1 from coho salmon (csPA), a cold water fish species, using circular dichroism, scanning calorimetry, hydrophobic probe fluorescence, limited proteolysis, chemical crosslinking and dynamic light scattering techniques. Apo-csPA is shown to be mainly monomeric protein with markedly disorganized secondary structure and lack of rigid tertiary structure. Examination of per-residue propensity for intrinsic disorder in the PA groups with either folded or unfolded apo-form using the average PONDR VSL2P profiles revealed that the N-terminal region that includes alpha-helix A, AB-loop and N-terminal half of alpha-helix B is predicted to be less ordered in PM with disordered apo-state. Application of the structural criteria developed for discrimination of disordered PM indicate that the latter comprise about 16-19% of all PM. We show that structural instability of apo-beta-PA serves as a hallmark of elevated calcium affinity of the protein. Therefore, the successful predictions of unstable apo-PAs might facilitate search for PM with maximal calcium affinity and possibly serving as calcium sensors.